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Customer-developed protocol: precise genome editing in mouse embryos via ex vivo injection of RNP complexes and long ssDNA
The development and refinement of CRISPR genome editing technology has provided researchers with a more efficient, streamlined, and faster approach for generating knockin mouse models as compared to classical gene targeting. The following protocol, generously provided by Dr. Benedikt Wefers, Laboratory Head at the German Center for Neurodegenerative Diseases (DZNE), employs ex vivo microinjection of Cas9-guide RNA (crRNA:tracrRNA duplex) ribonucleoproteins (RNPs) in tandem with long ssDNA templates for homology-directed repair (HDR) into mouse zygotes.
Benedikt Wefers, PhD
Using this approach, Dr. Wefers and colleagues were able to achieve efficient insertion of a fluorescent protein in frame to an endogenous gene, as demonstrated by the corresponding application data.
User-generated protocols are based on internal proof-of-concept experiments, customer collaborations, and published literature. In some cases, relevant results are discussed in our research news BioView blog articles. While we expect these protocols to be successful in your hands, they may not be fully reviewed or optimized. We encourage you to contact us or refer to the published literature for more information about these user-generated and -reported protocols.
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